AUTHOR=Popa Laura Ioana , Gheorghe Irina , Barbu Ilda Czobor , Surleac Marius , Paraschiv Simona , Măruţescu Luminiţa , Popa Marcela , Pîrcălăbioru Graţiela Grădişteanu , Talapan Daniela , Niţă Mihai , Streinu-Cercel Anca , Streinu-Cercel Adrian , Oţelea Dan , Chifiriuc Mariana Carmen 

TITLE=Multidrug Resistant Klebsiella pneumoniae ST101 Clone Survival Chain From Inpatients to Hospital Effluent After Chlorine Treatment

JOURNAL=Frontiers in Microbiology

VOLUME=11

YEAR=2021

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2020.610296

DOI=10.3389/fmicb.2020.610296

ISSN=1664-302X

ABSTRACT=<p>In this paper we describe the transmission of a multi-drug resistant <italic>Klebsiella pneumoniae</italic> ST101 clone from hospital to wastewater and its persistence after chlorine treatment. Water samples from influents and effluents of the sewage tank of an infectious diseases hospital and clinical strains collected from the intra-hospital infections, during a period of 10 days prior to wastewater sampling were analyzed. Antibiotic resistant <italic>K. pneumoniae</italic> strains from wastewaters were recovered on selective media. Based on antibiotic susceptibility profiles and PCR analyses of antibiotic resistance (AR) genetic background, as well as whole-genome sequencing (Illumina MiSeq) and subsequent bioinformatic analyses, 11 ST101 <italic>K. pneumoniae</italic> strains isolated from hospital wastewater influent, wastewater effluent and clinical sector were identified as clonally related. The SNP and core genome analyses pointed out that five strains were found to be closely related (with ≤18 SNPs and identical cgMLST profile). The strains belonging to this clone harbored multiple acquired AR genes [<italic>bla</italic><sub>CTX–M–</sub><sub>15</sub>, <italic>bla</italic><sub>OXA–</sub><sub>48</sub>, <italic>bla</italic><sub>OXA–</sub><sub>1</sub>, <italic>bla</italic><sub>SHV–</sub><sub>106</sub>, <italic>bla</italic><sub>TEM–</sub><sub>150</sub>, <italic>aac(3)-IIa</italic>, <italic>aac(6′)-Ib-cr</italic>, <italic>oqx</italic>A10, <italic>oqx</italic>B17, <italic>fos</italic>A, <italic>cat</italic>B3, <italic>dfr</italic>A14, <italic>tet</italic>(D)] and chromosomal mutations involved in AR (Δ<italic>mgrB</italic>, Δ<italic>ompK35</italic>, amino acid substitutions in GyrA Ser83Tyr, Asp87Asn, ParC Ser80Tyr). Twenty-nine virulence genes involved in iron acquisition, biofilm and pili formation, adherence, and the type six secretion system – T6SS-III were identified. Our study proves the transmission of MDR <italic>K. pneumoniae</italic> from hospital to the hospital effluent and its persistence after the chlorine treatment, raising the risk of surface water contamination and further dissemination to different components of the trophic chain, including humans.</p>